B6(Cg)-Rab32^tm1.1Ozg/J

Stock number: 040780
Product name: Rab32*S69R^flox
Research areas: Neurobiology Research, Research Tools

Description

Rab32*S69R^flox mice constitutively express a loxP-flanked mutant exon 1 harboring the S69R mutation of the Rab32 gene. The mouse S69R mutation corresponds to the human S71R mutation that is associated with Parkinson's disease. After Cre-mediated recombination, the floxed mutant exon 1 is excised resulting in a knock-out allele.

Detailed description

RAB32, member RAS oncogene family (Rab32) gene codes for an RAB GTPase, which plays a role in multiple cellular processes including endosomal transport, autophagy, generation of melanosomes, phagocytosis and inflammation. RAB32 has also been linked to regulation of LRRK2 kinase activity. RAB32 missense mutation S71R is a newly identified genetic risk factor for autosomal dominant Parkinson's disease; this variant leads to increased LRRK2 activity and symptoms such as tremors, postural instability, and motor fluctuations in patients harboring the S71R mutation. Rab32*S69R^flox mice constitutively express a loxP-flanked mutant exon 1 harboring the S69R mutation of the Rab32 gene. The mouse S69R mutation corresponds to the human S71R mutation. After Cre-mediated recombination, the floxed mutant exon 1 is excised resulting in a knock-out allele.

Heterozygous mice are viable and fertile with no overt phenotype. Homozygous mice are born in lower than expected Mendelian ratios (~12%), from heterozygous x heterozygous matings. The fertility of surviving homozygous mice has not been evaluated. After Cre-mediated recombination, the mutant exon 1 is excised in cre-expressing tissues, resulting in a knock-out (KO) allele. Homozygous KO mice are embryonic lethal. These Rab32*S69R^flox mice may be useful when studying familial Parkinson's disease.

Development

The Rab32*S69R^flox knock-in allele was designed to incorporate the S69R (AGC to AGG) mutation in exon 1 of the RAB32, member RAS oncogene family (Rab32) gene. The targeting construct consists of an initial loxP site, an F3-flanked puromycin cassette, mutant exon 1 harboring the S69R mutation, an FRT-flanked neomycin cassette, and second loxP site. The construct was electroporated into C57BL/6J-derived ICS embryonic stem (ES) cells and correctly targeted ES cells were injected into recipient blastocysts. Chimeric mice were crossed to C57BL/6J mice to confirm germline transmission. The resulting Rab32*S69R^flox mice were crossed to a FLP recombinase mouse line, on a C57BL/6 background (OzFlp), to remove the F3 and FRT-flanked puromycin/neomycin cassettes. Rab32*S69R^flox mice were then backcrossed to C57BL/6J for at least 1 generation by the donating laboratory prior to sending males to The Jackson Laboratory in 2025. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

In 2025, the heterozygous males sent to The Jackson Laboratory were used to freeze sperm. Samples from those males were analyzed via the miniMUGA 10,000 SNP panel. This showed more than 99.4% of the markers as C57BL/6J.

Allele

Symbol: Rab32^tm1.1Ozg
Name: targeted mutation 1.1, Ozgene
MGI accession ID: MGI:8250447
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: Rab32*S69R^flox
Marker symbol: Rab32
Marker name: RAB32, member RAS oncogene family
Marker synonyms: 2810011A17Rik; PARK26
Chromosome: 10
Strain of origin: C57BL/6J
Molecular note: The allele was designed to incorporate the S69R (AGC to AGG) mutation in exon 1 of the RAB32, a member of the RAS oncogene family (Rab32) gene. The targeting construct consists of an initial loxP site, a F3-flanked puromycin cassette, a mutant exon 1 harboring the S69R mutation, an FRT-flanked neomycin cassette, and a second loxP site. FLP recombinase removed the F3 and FRT-flanked puromycin/neomycin cassettes.
General note: C57BL/6J-derived ICS embryonic stem (ES) cells was used.